TRACE manual - Center for Statistical Genetics
Transcription
TRACE manual - Center for Statistical Genetics
TRACE : fasT and Robust Ancestry Coordinate Estimation version 1.01 Chaolong Wang1 Department of Biostatistics School of Public Health Harvard University April 9, 2015 The TRACE software2 is available at http://www.sph.umich.edu/csg/chaolong/LASER/ 1 Comments 2 This on the TRACE software can be sent to [email protected]. software is licensed under the GNU General Public License, version 3.0. Contents 1 Introduction 2 2 Getting started 2.1 Availability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.2 Installing TRACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.3 Running TRACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 3 3 3 3 Examples 3.1 Basic usage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.2 Parallel jobs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.3 Internal reference . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 4 6 6 4 Input files 4.1 parameterfile ( .conf ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.2 studyfile ( .geno), genotypefile ( .geno), and sitefile ( .site) . . . . . . . . . . . 4.3 coordinatefile ( .coord ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 7 7 8 5 Usage options 5.1 Main parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.2 Advanced parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3 Command line arguments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 9 10 11 6 Output files 6.1 .log and terminal outputs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2 .RefPC.coord and .RefPC.var . . . . . . . . . . . . . . . . . . . . . . . . . . 6.3 .ProPC.coord . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 13 13 14 7 Computational complexity 14 8 Version changes 8.1 Version 1.0 (May 19, 2014) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8.2 Version 1.01 (Apr 6, 2015) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 15 15 9 Acknowledgements 15 References 16 1 1 Introduction TRACE is a software program that uses SNP genotypes to trace individual ancestry by comparing to a set of reference individuals. The basic idea is to construct a reference ancestry map by applying principal components analysis (PCA) on genotypes of the reference individuals, and then to place the study samples one by one into this reference ancestry map. With an appropriate reference panel, the estimated coordinates of the study samples are informative on their ancestry background. To place each sample, we use a two-step approach involving PCA and projection Procrustes analysis (Gower and Dijksterhuis, 2004; Wang et al., 2010). The TRACE program follows the same framework as the LASER program, which we previously developed to estimate individual ancestry by directly analyzing shortgun reads from next generation sequencing without calling genotypes (Wang et al., 2014). Different from LASER, TRACE takes genotype data as the input, and thus can benefits studies when sequence read data are not available. TRACE can identify population structure in large cohorts. Compared to standard PCA, TRACE has several advantages in terms of robustness and computational efficiency. First, TRACE is robust to missing data in the study samples. Standard PCA requires high quality genotypes with low missing data rate for the study samples. Commonly used PCA software programs such as smartpca (Patterson et al., 2006) often impute missing data with mean values at the corresponding loci of the genotype matrix. Consequently, samples that have more missing data will shrink closer to the center of the PCA map. TRACE, in contrast, only requires low missingness in the reference individuals and can appropriately handle missing data in the study samples, which makes TRACE applicable to low quality data such as those obtained from ancient DNA samples (Skoglund et al., 2012). Second, TRACE is robust to the presence of close genetic relatedness between study samples. If PCA is applied direclty on all study samples, closely related individuals often appear as outliers, and might distort the overall PCA pattern. TRACE avoids this problem by analyzing each sample independently with the reference panel, which only requires the reference individuals to be unrelated with each other and with the study samples. For the same reason, TRACE is also robust to uneven sampling scheme and existence of genetic outliers in the study samples, which are known to have large impacts on PCA results (McVean, 2009; Lee et al., 2010). Last, the computational time of TRACE increases linearly with the number of study samples, so that TRACE can be much faster than standard PCA when the sample size is large. Details of the method and examples illustrating the aforementioned advantages of TRACE can be found in our paper entitled “ Improved ancestry estimation for both genotyping and sequencing data using projection Procrustes analysis and genotype imputation” (Wang et al., 2015). 2 2 Getting started 2.1 Availability The TRACE program is distributed as part of the LASER software package (Wang et al., 2014, 2015), which can be downloaded from the following webpage: http://www.sph.umich. edu/csg/chaolong/LASER/. In the package, we provide a pre-compiled executable for TRACE for Linux (64-bit) operation systems. This program is licensed under the GNU General Public License, version 3.0. A copy of the license is included in the package or can be found at http://www/gnu.org/licenses. Source code written in C++ is provided in the package. The TRACE program uses two external C++ libraries: the Armadillo Linear Algebra Library (http://arma.sourceforge. net) (Sanderson, 2010) and the GNU Scientific Library (http://www.gnu.org/software/ gsl). The Armadillo library requires two additional libraries: LAPACK and BLAS. Therefore, to compile from source code, you need to have these four libraries installed in your computer. Please cite the following papers when you use TRACE : 1. Wang et. al. (2014) Ancestry estimation and control of population stratification in sequence-based association studies. Nature Genetics, 46: 409-415. 2. Wang et al. (2015) Improved ancestry estimation for both genotyping and sequencing data using projection Procrustes analysis and genotype imputation. AJHG, (accepted). 2.2 Installing TRACE Open a terminal in the same directory as the .tar.gz file. Extract the file by typing tar -xzvf LASER-2.02.tar.gz in the terminal. This will create a new directory called LASER-2.02. This direcotry contains executables for both LASER (version 2.02) and TRACE (version 1.01), and other resource files. 2.3 Running TRACE Open a terminal and path to the directory that contains the executable TRACE. If you did not rename the directory after extracting the .tar.gz file, the directory will be LASER-2.02. Execute the program by typing ./trace -p parameterfile , in which -p is the command line flag specifying the parameter file and parameterfile is the name of the parameter file. If your parameterfile is not in the same directory, you must specify the whole path to the file. If the parameterfile is not specified, TRACE will search in the current directory for a parameterfile named “trace.conf”, and execute the program with parameter values specified in “trace.conf”. If this file does not exist, an empty template parameterfile named “trace.conf” will be created in the current directory. For more command line arguments, see Section 5.3. 3 3 Examples This section provides example usage of the TRACE program based on data in the folder named “example” (included in the download package). If you have questions when reading this section, please refer to the next few sections for detailed information about the input files (Section 4), usage options (Section 5), and output files (Section 6). The “example” folder contains genotype data across 9,608 SNPs on chromosome 22 for 938 individuals from the Human Genome Diversity Panel (HGDP, Li et al., 2008). The HGDP data are splited into two files named HGDP 700 chr22.geno and HGDP 238 chr22.geno, containing 700 and 238 individuals, respectively. The HGDP chr22.site file provides detailed information of the 9,608 SNP markers. The HGDP 238 chr22.RefPC.coord file contains PCA coordinates for the top 8 PCs based on genotypes in the HGDP 238 chr22.geno file. The folder also includes a parameterfile named “example.conf”, which specifies parameters for running TRACE on the example data. In the following examples, we will use the subset of 238 individuals to construct the reference PCA space and use TRACE to place the remaining 700 individuals into the reference PCA space. 3.1 Basic usage After decompressing the download package, enter the folder that contains the executable TRACE program. The following command will use parameter values provided in the example parameterfile (shown at the end of this section). ./trace -p ./example/example.conf The following command will change the number of PCs (DIM ) to 6 and the prefix of output file names (OUT PREFIX ) to “HGDP”, while the other parameters are defined by the example.conf file. ./trace -p ./example/example.conf -o HGDP -k 6 Results from TRACE will be output to the current working directory. The example parameterfile is similar to the one shown below. Each line specifies one parameter, followed by the parameter value (or followed by a “#” character if the parameter is undefined). Text after a “#” character in each line is treated as comment and will not be read by the program. # This is a parameter file for TRACE v1.01. # The entire line after a ‘#’ will be ignored. ###----Main Parameters----### STUDY_FILE ./example/HGDP_700_chr22.geno 4 # no default value GENO_FILE ./example/HGDP_238_chr22.geno # no default value COORD_FILE ./example/HGDP_238_chr22.RefPC.coord # no default value OUT_PREFIX test # default "trace" DIM 2 # default 2 DIM_HIGH 20 # default [20] MIN_LOCI 100 # default 100 ###----Advanced Parameters----### ALPHA # default 0.1 THRESHOLD # default 0.000001 REF_SIZE # default 200 FIRST_IND # default 1 LAST_IND # default [last sample in the STUDY_FILE] TRIM_PROP # default 0 MASK_PROP # default 0 EXCLUDE_LIST # no default value PROCRUSTES_SCALE # default 0 [include scaling as a free parameter] ###----Command line arguments----### # -p parameterfile (this file) # -s STUDY_FILE # -g GENO_FILE # -c COORD_FILE # -o OUT_PREFIX # -k DIM # -K DIM_HIGH # -l MIN_LOCI # -a ALPHA # -t THRESHOLD # -N REF_SIZE # -x FIRST_IND # -y LAST_IND # -M TRIM_PROP # -m MASK_PROP # -ex EXCLUDE_LIST # -rho PROCRUSTES_SCALE ###----End of file----### 5 3.2 Parallel jobs We currently do not implement multi-thread option to run parallel jobs. Because each study sample is analyzed independently, users can easily parallel the analyses by running multiple jobs simultaneously. The -x and -y flags provide a convenient way to specify a subset of samples to analyze in each job. For example, running the following commands will submit two jobs: the first job will analyze samples 1 to 350; and the second job will analyze samples 351 to 700 in the HGDP 700 chr22.seq file. ./trace -p ./example/example.conf -x 1 -y 350 -o results.1-350 & ./trace -p ./example/example.conf -x 351 -y 700 -o results.351-700 & Outputs from these two jobs will have different file name prefixes results.1-350 and results.351700 specified by the -o flag. We also recommend users to provide the coordinatefile when running multiple jobs using the same set of of reference individuals to save computational time by avoiding redundant calculation of the reference PCA coordinates in each job. 3.3 Internal reference In cases when external references are not available or when users want to focus on the study sample, TRACE provides an option to use a subset of individuals randomly from the study sample as an internal reference panel. The movitation here is to reduce computational cost when the sample size is too large for using standard PCA approaches. The size of the internal reference panel is defined by the parameter REF SIZE (-N). This option is only in effect when the parameter GENO FILE (-g) is undefined. For example, after commenting out the line for GENO FILE in the example.conf file, the following command will randomly select 300 individuals from the STUDY FILE as the reference panel, and run the TRACE analysis on the remaining 400 individuals. ./trace -p ./example/example.conf -N 300 Note that there is trade-off between computational efficiency and accuracy when using this internal reference option. Setting a smaller value for REF SIZE can reduce the computational time, but might lead to less accurate results compared to results based on standard PCA. In addition, the option for running multiple jobs in parallel using parameters FIRST IND (-x) and LAST IND (-y) is not available when using the internal reference option. 4 Input files In this section, we describe four input files that are taken by TRACE — the parameterfile, the studyfile, the genotypefile, and the coordinatefile. The genotypefile and the studyfile have 6 the same format. We also describe one additional file, the sitefile, which is associated with the the genotypefile and the studyfile. Note that the formats of the genotypefile, the sitefile, and the coordinatefile are the same as used by the LASER software (Wang et al., 2014). 4.1 parameterfile ( .conf ) The parameterfile contains all parameters required for running TRACE. The default parameterfile is “trace.conf”, which does not need to be explicitly specified in the command line (i.e. ./trace is equivalent to ./trace -p trace.conf). There are nine parameters in the parameterfile , including six main parameters and three advanced parameters. Each parameter is followed by its assigned value, separated by whitespaces. Text in the same line after a ‘#’ character is treated as comment and will not be read. For example, the following parameter specifications are equivalent in setting the parameter DIM equal to 4: DIM 4 DIM 4 # Number of PCs to compute DIM 4 # Other comments If the user does not assign a value to a parameter in the parameterfile, this parameter must be followed by a ‘#’ character (even without comments) to avoid unexpected errors in assigning other parameter values. An example parameterfile is provided in Section 3.1. To generate an empty template parameterfile, run TRACE when the default parameterfile does not exist and without any command line arguments. Three parameters do not have default values, among which two parameters (GENO FILE and STUDY FILE ) need to be explicitly defined by the users when in use, either in the parameterfile or in the command line (see Section 5.3), and one parameter (COORD FILE ) is optional. The other six parameters do not need to be explicitly defined unless the user wants to use settings different from the default. Please refer to Section 5 for more information on these parameters. 4.2 studyfile ( .geno), genotypefile ( .geno), and sitefile ( .site) The studyfile and the genotypefile contain genotype data for the study sample and the reference panel, respectively. TRACE does not require the genotypefile and the studyfile to contain same set of loci in the same order. Detail information of the loci should be provided in the sitefile’s. TRACE will automatically extract loci shared by the studyfile and the genotypefile for downstream analyses. Each line in the genotypefile/studyfile represents genotype data of one individual. The first two columns represent population IDs and individual IDs, respectively. Starting from the third column, each column represent a locus. We only consider bi-allelic SNP markers. 7 To be consistent with the sequence data, genotypes should be given on the forward strand. Genotypes are coded by 0, 1, or 2, representing copies of the reference allele at a locus in one diploid individual. Missing data are coded by -9. TRACE can also be applied to multi-ploidy organisms. In general, genotypes should be coded by 0, 1, ..., K for K-ploidy organisms. Columns in the genotypefile/studyfile are tab-delimited. An example is provided below: POP_1 POP_1 POP_2 POP_3 ... IND_1 IND_2 IND_3 IND_4 ... 2 2 0 1 ... 0 -9 0 2 ... 1 2 -9 1 ... ... ... ... ... ... Information on each locus, including chromosome number, genomic position, SNP ID, reference allele, and alternative allele, is listed in a separate sitefile. The reference allele and the alternative allele should be given on the forward strand. The first row of the sitefile is the header line. Starting from the second line, each line represents one locus. Columns in the sitefile are tab-delimited. An example sitefile is provided below: CHR 1 1 1 ... POS 752566 768448 1005806 ... ID rs3094315 rs12562034 rs3934834 ... REF G G C ... ALT A A T ... To run TRACE, a “.site” file is required for each “.geno” file. Users can convert their genotype data from VCF format to our “.geno” and “.site” format using a program called vcf2geno. This program is available from the LASER software package (http://www.sph. umich.edu/csg/chaolong/LASER/). The command line for running vcf2geno is ./vcf2geno --inVcf filename.vcf --out output which will generate a genotypefile named “output.geno” and a sitefile named “output.site”. 4.3 coordinatefile ( .coord ) The coordinatefile contains PCA coordinates of the reference individuals. The first line is the header line. Starting from the second line, each line represent one individual. The first two columns correspond to population IDs and individual IDs respectively, and the following K columns represent the top K principal components (PCs). The order of the reference individuals must be the same as in the genotypefile. The coordinatefile is required to be tab-delimited. Below is an illustration of the format of the coordinatefile: 8 popID POP_1 POP_1 POP_2 POP_3 ... indivID IND_1 IND_2 IND_3 IND_4 ... PC1 -3.5 -2.2 7.8 1.6 ... PC2 0.2 4.5 -0.8 -3.8 ... PC3 0.7 0.8 -1.0 -0.4 ... ... ... ... ... ... ... If the coordinatefile is not provided, TRACE will automatically compute the reference coordinates based on the genotypefile, and the results will be output. We recommend users to prepare a coordinatefile as input for TRACE when submitting multiple jobs using the the same reference panel (so that the same computation will not be repeated for every job). 5 Usage options TRACE has 16 parameters that users can set in the parameterfile, including 7 main parameters that are required for running TRACE and 9 advanced parameters for some special options. Among the 7 main parameters, 3 are parameters regarding the input data files and need to be explicitly defined when in use. The other 4 main parameters and the 9 advanced parameters have default values. In addition, TRACE takes 17 command line arguments, which are described in Section 5.3. 5.1 Main parameters STUDY FILE (string) The name of the studyfile. If the file is not in the same directory as TRACE, the whole path must be specified. This parameter must be explicitly defined. GENO FILE (string) The name of the genotypefile. If the file is not in the same directory as TRACE, the whole path must be specified. This parameter must be explicitly defined unless using the internal reference option. COORD FILE (string) The name of the coordinatefile. If the file is not in the same directory as TRACE, the whole path must be specified. This parameter is optional. If undefined, TRACE will automatically compute the reference coordinates based on the genotypefile. OUT PREFIX (string) The prefix that will be added to the file names of outputting results. A path can be specified to output results to a different directory. The default value is “TRACE ”. 9 DIM (int) The number of PCs to compute (must be a positive integer). This number must be smaller than the number of individuals and the number of loci in the genotypefile, and cannot be greater than the number of PCs in the coordinatefile if a coordinatefile is provided. The default value is 2. DIM HIGH (int) Dimension of the sample-specific PCA map to project from (must be a positive integer). This number must be smaller than the number of individuals and the number of loci in the genotypefile, and cannot be smaller than DIM. TRACE will project each study sample from a DIM HIGH dimensional PC space to the DIM dimensional reference ancestry map. If set to 0, the program will use the number of significant PCs based on TracyWidom tests for each sample. The default value is 20. MIN LOCI (int) The minimum number of non-missing loci required for an individual in the study sample to be analyzed (must be a positive integer). If the number of non-missing loci is smaller than MIN LOCI, the individual will not be analyzed and results for this individual are output as “NA”. The default value is 100. 5.2 Advanced parameters ALPHA (double) Significance level in Tracy-Widom tests to determine the number of informative PCs, or DIM HIGH, in the sample-specific PCA (must be a number between 0 and 1). This parameter is effective only if DIM HIGH is undefined or set to 0. The default value is 0.1. THRESHOLD (double) Convergence criterion of the projection Procrustes analysis (must be a positive number). The default value is 0.000001. REF SIZE (int) The number of individuals to be randomly selected from the study sample as an internal reference panel (must be a positive integer). This number cannot be greater than the number of individuals in the studyfile. This parameter will be in effect only if GENO FILE is undefined (i.e., an external reference panel is not provided). The default value is 200. FIRST IND (int) The index of the first individual in the study sample to analyze (must be a positive integer). This number cannot be greater than the number of individuals in the studyfile. Individuals that have indices smaller than FIRST IND will be skipped. This parameter will be in effect only if GENO FILE is defined (i.e., an external reference panel is provided). The default value is 1. LAST IND (int) The index of the last individual in the study sample to analyze (must 10 be a positive integer). This number cannot be greater than the number of samples in the studyfile or smaller than FIRST IND. Individuals that have indices greater than LAST IND will be skipped. This parameter will be in effect only if GENO FILE is defined (i.e., an external reference panel is provided). The default value is the number of samples in the studyfile. TRIM PROP (double) Proportion of randomly selected loci to exclude from the analysis for all samples (must be a number between 0 and 1). This option is useful when the data size is too big such that memory limit becomes an issue. The default value is 0. MASK PROP (double) Proportion of loci in a study sample that will be randomly set to missing (must be a number between 0 and 1). This option is useful when testing the robustness to missing data or examing the minimal number of markers requested in a sample in order to have satisfying performance given a reference panel. The default value is 0. EXCLUDE LIST (string) This parameter specifies the file name of a list of SNPs to exclude from the analysis. Each line of the file is a SNP ID and no header is required. If the file is not in the same directory as TRACE, the whole path must be specified. This parameter do not have default value. PROCRUSTES SCALE (int) This parameter specifies how to scale coordinates in the Procrustes analysis (must be 0 or 1). When set to 0, the Procrustes analysis will include the scaling factor as a free parameter and estimates its value to minimize the sum of squared Euclidean distances between two sets of coordinates in the analysis. When set to 1, the Procrustes analysis will fix the scaling factor so that two sets of coordinates will have the same total variance (i.e. Procrustes analysis only search for rotation, reflection and translation to minimize the sum of squared Euclidean distances). The default value is 0. 5.3 Command line arguments The command line flags provide the user an option to enter information from the command line. All command line arguments will overwrite values specified in the parameterfile. If a parameter is specified with an invalid value in the parameterfile but a valid value in the command line, the program will return a warning message and still execute correctly by taking the value from the command line. However, if a parameter value in the command line is not valid, the program will exit with an error message. If a command line flag is specified, it must be followed by a space and then the parameter value. Different command line flags can appear in any order. If the same command line flag is defined more than once, only the last value will be taken. For example, the following command lines are equivalent and will change the value 11 of the parameter DIM, for which the command line flag is -k, to be 4 while using the other parameters defined in the parameterfile named “my parameterfile”. ./trace -p my_parameterfile -k 4 ./trace -k 4 -p my_parameterfile ./trace -k 3 -p my_parameterfile -k 4 Most command line arguments are optional except for the parameterfile, for which the command line flag is -p. A list of all command line flags is provided below. -p This flag defines the parameterfile. If the parameterfile is not in the current directory, a whole path to the file must be specified. This parameter can only be defined using the command line. If undefined, the program will use the default parameterfile named “trace.conf” in the current directory. If this file does not exist, an empty template parameterfile named “trace.conf” will be created in the current directory, and the program will then exit with an error message. -s Change the parameter value of STUDY FILE. -g Change the parameter value of GENO FILE. -c Change the parameter value of COORD FILE. -o Change the parameter value of OUT PREFIX (useful when running parallel jobs). -k Change the parameter value of DIM. -K Change the parameter value of DIM HIGH. -l Change the parameter value of MIN LOCI. -a Change the parameter value of ALPHA. -t Change the parameter value of THRESHOLD. -N Change the parameter value of REF SIZE. -x Change the parameter value of FIRST IND (useful when running parallel jobs). 12 -y Change the parameter value of LAST IND (useful when running parallel jobs). -M Change the parameter value of TRIM PROP (useful when memory is limited). -m Change the parameter value of MASK PROP. -ex Change the parameter value of EXCLUDE LIST. -rho Change the parameter value of PROCRUSTES SCALE. 6 Output files All output files will be saved in the current directory unless the path to a different directory is given in the parameter value of OUT PREFIX. All output file names will start with the parameter value of OUT PREFIX. These files are described below. 6.1 .log and terminal outputs The terminal outputs are used to monitor and record the progress when running TRACE. It starts with all parameter values used in the execution of TRACE, and reports the progress of the program step by step. The log file is identical to the terminal outputs. 6.2 .RefPC.coord and .RefPC.var When COORD FILE is not defined, TRACE will perform PCA on the reference genotype data given by the genotypefile. Results of the top k PCs, where k is defined by the parameter DIM, are output to two files named OUT PREFIX.RefPC.coord and OUT PREFIX.RefPC.var. The RefPC.coord file records the PCA coordinates of the reference individuals. The first line in this file is a header line. Starting from the second line, each line represents one individual. The first two columns are population ID and individual ID, respectively. The remaining columns correspond to the top k PCs. This file is tab-delimited. The format of this file is exactly the same as the coordinatefile (Section 4.3), so that this file can be directly used as the input file for TRACE. The RefPC.var file records the proportion of variance explained by each PC. The first line in this file is a header line. Starting from the second line, each line represents one PC. The first column is the PC index and the second column is the percentage of variance explained by each PC. Only results for the top k PCs are output. This file is tab-delimited. 13 6.3 .ProPC.coord This file contains the estimated PCA (or “Procrustean PCA”) coordinates of the study sample. The first line is a header line. Starting from the second line, each line represents one study sample. The first column is population ID, and the second column is individual ID. The third column reports the number of nonmissing loci used in the analysis. The fourth column reports values of DIM HIGH, dimension of the sample-specific PCA map used in projection Procrustes analysis. The fifth column reports the Procrustes similarity score between each sample-specific PCA map (after being projected to the DIM dimensional space) and the original DIM dimensional reference PCA map. Starting from the sixth column, each column represents coordinates of one PC (up to the kth PC, where k is defined by DIM ). Columns in this file are tab-delimited. When using the internal reference option, coordinates for the individuals that are selected as the internal reference will also be included in this file. The values of DIM HIGH (the fourth column) and the Procrustes similarity scores (the fifth column) for these individuals will be listed as “NA”. 7 Computational complexity TRACE examines one study individual at a time. Therefore, the computational costs increase linearly with the number of individuals to be analyzed. We can easily run the analyses in parallel by submitting multiple jobs to analyze different subsets of the study sample (using command line flags -x and -y). The cost for analyzing each individual depends on the number of individuals, N , and the number of loci, L, in the reference panel, and the proportion of missing data, m, in the study individual. We first calculate the N × N pairwise similarity matrix of the reference panel, for which the computational cost is O(N 2 L). This computation is only performed once and will be repeatedly used in analyzing each individual. Roughly, we expect computational cost of PCA for each study individual to be O(N L + N 2 Lm + N 3 ), in which N L is the time required for comuputing the extra row (and column) for the study individual in the similarity matrix, N 2 Lm is for adjustment of the similarity matrix to account for missing data in the study individual, and N 3 is for eigen decomposition on the similarity matrix. The computational cost of projection Procrustes analysis is approximately O[q(N K + K 3 )], where q is the number of iterations required for projection Procrustes analysis to converge and K is the value of DIM HIGH. When K is not big, the analysis often converges within a few iterations, and the cost for projection Procrustes analysis is negligible compared to the cost for PCA. Overall, the computational cost for TRACE on a study sample of n individuals is approximately O[N 2 L + n(N L + N 2 Lm ¯ + N 3 + qN K + qK 3 )]. If we ignore the cost for 14 projection Procrustes analysis and suppose the missing data rate is small, the computational cost for TRACE can be approximated as O[N 2 L + n(N L + N 3 )]. In comparison, the cost for a standard PCA on n individuals is approximately O(n2 L + n3 ). 8 Version changes Changes from previous versions of the TRACE software are noted here. 8.1 Version 1.0 (May 19, 2014) - Initial release of the TRACE software. 8.2 Version 1.01 (Apr 6, 2015) - Added a new parameter PROCRUSTES SCALE to allow scaling two sets of coordinates to have the same variance in the Procrustes analysis. - Fixed a minor bug to in the projection Procrustes anlaysis (convergence issue, no impact on the results). 9 Acknowledgements I would like to thank Conrad Sanderson at the National ICT Australia for his help on using the Armadillo library. References Gower, J. C., and G. B. Dijksterhuis, 2004 Press. Procrustes Problems. Oxford University Lee, A. B., D. Luca, L. Klei, B. Devlin and K. Roeder, 2010 ancestry using spectral graph theory. Genet. Epidemiol. 34: 51–59. Discovering genetic Li, J. Z., D. M. Absher, H. Tang, A. M. Southwick, A. M. Casto, S. Ramachandran, H. M. 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